Biochemical, pathological and molecular characterisation of Phomopsis vexans: A causative of leaf blight and fruit rot in brinjal.

The pathogen Phomopsis vexans causes leaf blight, fruit rot, and damping off in brinjal plants, all of which are extremely detrimental. The pathogen affects host plant photosynthetic efficiency and fruit quantity and quality. An appreciation of the pathogenicity of P. vexans is essential for the effective control of infections in the field. Consequently, the goal of this study was to characterise P. vexans in terms of their biochemistry, molecular diversity, and pathogenicity. In terms of cellulase (97.7 U), catalase (12.2 U), and ascorbate peroxidase (147.3 U) activity, isolate PV1 performed best, followed by PV5 (CL-97.0 U, CAT-11.1 U and APX-144.4 U), and PV8 (CL-88.8 U, CAT-9.8 U and APX-141.9 U). In a greenhouse pathogenicity test, isolate PV1 had the highest incidence (97%) and severity (88.6%) of disease, whereas isolate PV6 showed the lowest incidence (57.2%) and severity (70%) of disease. The biochemical enzyme activity of P. vexans corresponds well with its greenhouse pathogenicity results, and its combination can be exploited to identify pathogenic P. vexans isolates. Using RAPD and ISSR primers, molecular characterisation indicated genetic diversity but could not distinguish isolates by geographical origin or pathogenicity. The pathogen P. vexans was verified by ITS1 and ITS4 molecular analysis, and the sequences were subsequently deposited in the NCBI database. In conclusion, the enzyme activity relevant to pathogenicity (CL, CAT and APX) in conjunction with the invivo pathogenicity assay might be utilised to differentiate between pathogenic (virulent) and non-pathogenic (avirulent) P. vexans isolates and develop suitable disease management strategies.

Development of Wash-Durable Antimicrobial Cotton Fabrics by In Situ Green Synthesis of Silver Nanoparticles and Investigation of Their Antimicrobial Efficacy against Drug-Resistant Bacteria.

An environment friendly and wash-durable silver nanoparticle treatment of cotton fabrics was carried out by in situ reduction of silver nitrate using Azadirachta indica leaf extract. The wash durability of the silver nanoparticles treatment on the cotton fabric was improved by pretreating the fabrics by mercerization and by adopting hydrothermal conditions of 120 °C temperature and 15 psi pressure for the in situ synthesis. The silver nanoparticle treated fabrics were characterized using scanning electron microscopy, colorimetric analysis and inductively coupled plasma mass spectroscopy. The coating of silver nanoparticles was seen to be dense and uniform in the scanning electron micrographs of the treated fabrics. An evaluation of the antibacterial efficacy of the silver nanoparticle treated fabric against antibiotic-resistant Gram-positive and Gram-negative strains was carried out. The antibacterial efficacy was found to be the highest against Bacillus licheniformis, showing 93.3% inhibition, whereas it was moderate against Klebsiella pneumoniae (20%) and Escherichia coli (10%). The transmittance data of a UV spectrophotometer (290-400nm) was used for measuring the UV protection factor of the silver nanoparticle treated fabrics. All the silver nanoparticle treated fabrics showed good antimicrobial and UV protection activity. The treatment was also seen to be durable against repeated laundering. This paper contributes the first report on a novel green synthesis approach integrating mercerization of cotton fabrics and in situ synthesis of nanoparticles under hydrothermal conditions using Azadirachta indica leaf extract for improved wash durability of the multifunctional fabric.

Trichovariability in rhizosphere soil samples and their biocontrol potential against downy mildew pathogen in pearl millet.

The present work is aimed to examine the genetic variability and the distribution pattern of beneficial Trichoderma spp. isolated from rhizosphere samples and their mode of action in improving the plant health. A total of 131 suspected fungi were isolated from the rhizospheric soil and 91 isolates were confirmed as Trichoderma spp. T. asperellum and T. harzianum were found high in the frequency of occurrence. Genetic diversity analysis using RAPD and ISSR revealed the diverse distribution pattern of Trichoderma spp. indicating their capability to adapt to broad agroclimatic conditions. Analysis of genetic diversity using molecular markers revealed intra-species diversity of isolated Trichoderma spp. The frequency of pearl millet (PM) root colonization by Trichoderma spp. was found to be 100%. However, they showed varied results for indole acetic acid, siderophore, phosphate solubilization, ß-1,3-glucanase, chitinase, cellulase, lipase, and protease activity. Downy mildew disease protection studies revealed a strong involvement of Trichoderma spp. in direct suppression of the pathogen (mean 37.41) in the rhizosphere followed by inducing systemic resistance. Our findings highlights the probable distribution and diversity profile of Trichoderma spp. as well as narrate the possible utilization of Trichoderma spp. as microbial fungicides in PM cultivation across different agroclimatic zones of India.

In silico studies evidenced the role of structurally diverse plant secondary metabolites in reducing SARS-CoV-2 pathogenesis.

Plants are endowed with a large pool of structurally diverse small molecules known as secondary metabolites. The present study aims to virtually screen these plant secondary metabolites (PSM) for their possible anti-SARS-CoV-2 properties targeting four proteins/ enzymes which govern viral pathogenesis. Results of molecular docking with 4,704 ligands against four target proteins, and data analysis revealed a unique pattern of structurally similar PSM interacting with the target proteins. Among the top-ranked PSM which recorded lower binding energy (BE), > 50% were triterpenoids which interacted strongly with viral spike protein-receptor binding domain, > 32% molecules which showed better interaction with the active site of human transmembrane serine protease were belongs to flavonoids and their glycosides, > 16% of flavonol glycosides and > 16% anthocyanidins recorded lower BE against active site of viral main protease and > 13% flavonol glycoside strongly interacted with active site of viral RNA-dependent RNA polymerase. The primary concern about these PSM is their bioavailability. However, several PSM recorded higher bioavailability score and found fulfilling most of the drug-likeness characters as per Lipinski's rule (Coagulin K, Kamalachalcone C, Ginkgetin, Isoginkgetin, 3,3'-Biplumbagin, Chrysophanein, Aromoline, etc.). Natural occurrence, bio-transformation, bioavailability of selected PSM and their interaction with the target site of selected proteins were discussed in detail. Present study provides a platform for researchers to explore the possible use of selected PSM to prevent/ cure the COVID-19 by subjecting them for thorough in vitro and in vivo evaluation for the capabilities to interfering with the process of viral host cell recognition, entry and replication.

Opportunities and challenges in managing antibiotic resistance in bacteria using plant secondary metabolites.

Development of antibiotic resistance (ABR) in bacteria and its multidimensional spread is an emerging global threat that needs immediate attention. Extensive antibiotics (AB) usage results in development of ABR in bacteria by target modification, production of AB degrading enzymes, porin modifications, efflux pumps overexpression, etc. To counter this, apart from strict regulation of AB use and behavioural changes, research and development (R&D) of newer antimicrobials are in place. One such emerging approach to combat ABR is the use of structurally and functionally diverse plant secondary metabolites (PSMs) in combination with the conventional AB. Either the PSMs are themselves antimicrobial or they potentiate the activity of the AB through a range of mechanisms. However, their use is lagging due to poor knowledge of mode of action, structure-activity relationships, pharmacokinetics, etc. This review paper discussed the opportunities and challenges in managing ABR using PSMs. Mechanisms of ABR development in bacteria and current strategies to counter them were studied and the areas where PSMs can play an important role were highlighted. The use of PSMs, both as an anti-resistance and anti-virulence agent in combination therapy to counter multi-drug resistance along with their mechanisms of action, has been discussed in detail. The difficulties in the commercialisation of PSMs and strategies to overcome them along with future priority areas of research have also been given. Following the given R&D path will definitely help in better understanding and utilising the full potential of PSMs in solving the problem of antimicrobial resistance (AMR).

Elicitation of Novel Trichogenic-Lipid Nanoemulsion Signaling Resistance Against Pearl Millet Downy Mildew Disease.

Nanoemulsion was formulated from membrane lipids of Trichoderma spp. with the non-ionic surfactant Tween 80 by the ultrasonic emulsification method. Nanoemulsion with a droplet diameter of 5 to 51 nm was obtained. The possible effects of membrane lipid nanoemulsion on pearl millet (PM) seed growth parameters and elicitation of downy mildew (DM) disease resistance in PM was analyzed to develop an eco-friendly disease management strategy. Seed priming with nanoemulsion illustrates significant protection and elevated levels of early defense gene expression. Lipid profiling of Trichoderma spp. reveals the presence of oleic acid as a major fatty acid molecule. The prominent molecule in the purified lipid fraction of T. brevicompactum (UP-91) responsible for the elicitation of induction of systemic resistance in PM host against DM pathogen was predicted as (E)-N-(1, 3-dihydroxyoctadec-4-en-2yl) acetamide. The results suggest that protection offered by the novel nanoemulsion formulation is systemic in nature and durable and offers a newer sustainable approach to manage biotrophic oomycetous pathogen.

Characterization of non-aflatoxigenic strains of Aspergillus flavus as potential biocontrol agent for the management of aflatoxin contamination in groundnut.

In the present study, nine non-aflatoxigenic Aspergillus flavus strains were evaluated for their potential to reduce A. flavus infection and aflatoxin contamination in groundnut. Genetic characterization of these strains revealed six different deletion patterns (A-F) for thirteen examined genes from the aflatoxin biosynthesis pathway. Strain AFGS5 recorded maximum number of gene deletion (F) which included 12 out of 13 tested genes. Our findings indicated that aflR was the most frequently absent gene among the observed deletion patterns. A dendrogram inferred from combining random amplified polymorphic DNA and microsatellite data showed three of the non-aflatoxigenic strains segregating from other sampled isolates (aflatoxigenic and non-aflatoxigenic) tested. Greenhouse experiments, involving non-aflatoxigenic strains as biocontrol agents resulted in two strains, AFGS5 and AFGS12, which significantly reduced the population of aflatoxigenic fungi and the level of total aflatoxins in the rhizosphere/geocarposphere of soil samples as well as in groundnut seeds. Based on our findings, the use of these native non-aflatoxigenic strains; AFGS5 or AFGS12 in particular, as biopesticides could offer efficacious mitigation of aflatoxin contamination.